manumycin and Colorectal-Neoplasms

The use of natural products as chemotherapeutic agents and tools for manipulation of apoptosis represent an attractive therapeutic concept. In this study, we investigated the anticancer activities of a combination of two natural compounds with different origin, hypericin (plant product) in its photoactive state and Manumycin A (yeast product) and explored the underlying mechanisms of their pro-apoptotic action using an oxaliplatin-resistant variant of human colon adenocarcinoma cell line HT-29-OxR as the experimental model. CCK-8 assay was performed to evaluate the cytotoxicity of the drugs. CalcuSyn software was used to identify the type of interaction between the two agents. BrdU incorporation assay and colony forming assay were performed to study the short- and long-term proliferation of cells. To evaluate the ability of the drug combination to induce apoptosis, PARP p85 fragment was detected using the ELISA method. Changes in apoptosis-related proteins were examined by immunoassays. Our results showed that a synergistic combination of photoactive hypericin and Manumycin A decreased viability, inhibited both short- and long-term cell proliferation, decreased levels of IAPs proteins (cIAP1, cIAP2, XIAP and survivin), induced an apoptotic PARP cleavage associated with decline in procaspase-3 level, promoted phagocytosis of cancer cells, and restored chemosensitivity to oxaliplatin.

Topics: Anthracenes; Antineoplastic Agents; Apoptosis Regulatory Proteins; Cell Line; Cell Proliferation; Cell Survival; Colorectal Neoplasms; Drug Resistance, Neoplasm; Drug Synergism; Humans; Light; Macrophages; Oxaliplatin; Perylene; Phagocytosis; Polyenes; Polyunsaturated Alkamides

The chemopreventive effect of RAS inhibitors on colorectal cancer is unknown. Because aberrant crypt foci (ACF), earliest preneoplastic lesions, are highly positive for K-RAS mutation, RAS inhibitors are likely to be effective for chemoprevention. Therefore, in the present study, the suppressive effect of a RAS inhibitor, manumycin A, on ACF formation in an azoxymethane (AOM)-induced rat colorectal carcinogenesis model was investigated.. Rats injected with AOM were administered manumycin A (30 mg/kg) subcutaneously thrice weekly for 8 weeks or for 4 weeks (latter half), sacrificed at 8 weeks, and examined for ACF in the colorectum. Phosphorylated ERK and Ki-67 expression was evaluated by immunohistochemistry. Apoptosis was assessed by TUNEL staining.. The mean number of ACF in the 8-week manumycin A group (72.9 ± 20.1) was significantly lower than in the vehicle group (155.6 ± 56.7, P < 0.01), and it was significantly lower even in the 4-week manumycin A group than in the vehicle group (92.2 ± 13.0 vs 222.3 ± 83.3, P < 0.01). The positive rate for phosphorylated ERK in the manumycin A group (13.5 ± 19.2%) was significantly lower than in the vehicle group (50.2 ± 19.8%, P < 0.01). The positive rate for Ki-67 in the manumycin A group (2.2 ± 3.4%) was significantly lower than in the vehicle group (14.7 ± 8.2%, P < 0.01). There were significantly more terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling-positive cells in tissue samples from the manumycin A group versus the vehicle group (8.6 ± 9.7% vs 2.9 ± 2.0%, P < 0.05).. Manumycin A suppressed ACF formation in the AOM-induced colorectal carcinogenesis model, demonstrating that RAS inhibitors may be very effective for chemoprevention of colorectal cancers.

Topics: Aberrant Crypt Foci; Animals; Apoptosis; Azoxymethane; Colorectal Neoplasms; Disease Models, Animal; Enzyme Inhibitors; Extracellular Signal-Regulated MAP Kinases; Farnesyltranstransferase; Genes, ras; Injections, Subcutaneous; Ki-67 Antigen; Mutation; Phosphorylation; Polyenes; Polyunsaturated Alkamides; ras Proteins; Rats; Rats, Inbred F344

Recent studies have suggested that aberrant K-ras signaling is responsible for triggering immunological responses and inflammation-driven tumorigenesis. Interleukins IL-17, IL-22, and IL-23 have been reported in various types of malignancies, but the exact mechanistic role of these molecules remains to be elucidated. Given the role of K-ras and the involvement of interleukins in colorectal tumorigenesis, research efforts are reported for the first time, showing that differentially expressed interleukin IL-17, IL-22, and IL-23 levels are associated with K-ras in a stage-specific fashion along colorectal cancer progression. Specifically, a) the effect of K-ras signaling was investigated in the overall expression of interleukins in patients with colorectal cancer and healthy controls, and b) an association was established between mutant K-ras and cytokines GM-CSF and IFN-γ. The results indicate that specific interleukins are differentially expressed in K-ras positive patients and the use of K-ras inhibitor Manumycin A decreases both interleukin levels and apoptosis in Caco-2 cells by inhibiting cell viability. Finally, inflammation-driven GM-CSF and IFN-γ levels are modulated through interleukin expression in tumor patients, with interleukin expression in the intestinal lumen and cancerous tissue mediated by aberrant K-ras signaling. Collectively, the findings a) indicate that interleukin expression is influenced by ras signaling and specific interleukins play an oncogenic promoter role in colorectal cancer, highlighting the molecular link between inflammation and tumorigenesis, and b) accentuate the interwoven molecular correlations as leads to new therapeutic approaches in the future.

Topics: Adult; Aged; Aged, 80 and over; Apoptosis; Caco-2 Cells; Cell Survival; Colorectal Neoplasms; Disease Progression; Dose-Response Relationship, Drug; Female; Gene Expression Profiling; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-17; Interleukin-22; Interleukin-23; Interleukins; Male; Middle Aged; Mutation; Neoplasm Staging; Polyenes; Polyunsaturated Alkamides; Proto-Oncogene Proteins; Proto-Oncogene Proteins p21(ras); ras Proteins; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction